TransIT -Prostate Transfection Kit
|
|
- Sibyl Houston
- 6 years ago
- Views:
Transcription
1 INTRODUCTION TransIT -Prostate Transfection Kit is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in Prostate cell types. Generally, prostate cell types have been moderately difficult to transfect, yet have remained a prevalent cell line in the field of cancer research. TransIT-Prostate Transfection Kit contains two components, namely: TransIT -Prostate Reagent and the Prostate Boost Reagent. This kit provides all the attributes of the trusted TransIT series of transfection reagents: high transfection efficiency, low toxicity, serum compatibility, simplicity of use and reproducibility. Transfection using TransIT-Prostate Transfection Kit does not require medium changes and can be carried out in serum-containing medium. This kit is suitable for both transient and stable transfection. SPECIFICATIONS Storage Product Guarantee MATERIALS Materials Supplied Store both TransIT-Prostate Reagent and Prostate Boost Reagent at -20 C. Before each use, warm to room temperature and vortex gently. 6 months from the date of purchase, when properly stored and handled. Warm TransIT-Prostate and Prostate Boost Reagent to room temperature and vortex gently before each use. TransIT-Prostate Transfection Kit is supplied in one of the following formats. Product No. Volume of TransIT-Prostate Reagent Volume of Prostate Boost Reagent MIR ml ml MIR ml ml MIR ml ml MIR ml ml Materials required, but not supplied Cultured Prostate cells Appropriate cell culture medium Purified DNA Serum-free medium (e.g. Opti-MEM I Reduced-Serum Medium) Sterile tube for transfection complex preparation Micropipets Reporter assay as required For Research Use Only.
2 BEFORE YOU START: Important Tips for Optimal DNA Transfection Optimize reaction conditions for each Prostate cell subtype to ensure successful transfections. The suggestions below yield high efficiency DNA transfection using the TransIT-Prostate Transfection Kit. Table 1 on Page 3 presents recommended starting conditions depending on culture vessel size. Cell density (% confluence) at transfection. The recommended cell density for Prostate cell types at transfection is 50 70% confluence. Determine the optimal cell density for each Prostate cell type in order to maximize transfection efficiency. Divide the cells hours before transfection to ensure that the cells are actively dividing and reach the appropriate cell density at the time of transfection. DNA purity. Use highly purified, sterile, and contaminant-free DNA for transfection. Plasmid DNA preps that are endotoxin-free and have A 260/280 absorbance ratio of are desirable. DNA prepared using miniprep kits is not recommended as it may contain high levels of endotoxin. We recommend using MiraCLEAN Endotoxin Removal Kit (MIR 5900) to remove any traces of endotoxin from your DNA preparation. TransIT-Prostate Reagent:DNA ratio. As a starting point, use 2 µl of TransIT-Prostate Reagent per 1 µg of DNA. The optimal TransIT-Prostate Reagent to DNA ratio can be determined by titrating the reagent from 2 5 µl per µg of DNA. Please refer to Table 1 on Page 3 for recommended starting conditions. Prostate Boost Reagent:DNA ratio. Different Prostate Boost Reagent amounts may be required depending on the cell culture and experimental conditions. The optimal Prostate Boost Reagent:DNA ratio should be determined by titrating the reagent from 0 5 µl per µg of DNA, e.g., we find increased transfection efficiencies on LNCaP cells when 2 5 µl of Prostate Boost Reagent is added per 1 µg of DNA, whereas 0 2 µl of Prostate Boost Reagent per 1 µg of DNA is optimal for DU 145 cells. Refer to Table 1 on Page 3 for recommended starting conditions. Complex formation conditions. Prepare TransIT-Prostate:Prostate Boost:DNA complexes in serum-free growth medium. Mirus recommends Opti-MEM I Reduced-Serum Medium. Cell culture conditions: Culture cells in the appropriate medium, with or without serum. There is no need to perform a medium change to remove the transfection complexes. TransIT-Prostate Transfection Kit yields improved efficiencies when transfections are performed in complete growth medium (instead of serum-free medium) without a posttransfection medium change. Presence of antibiotics: Antibiotics will inhibit transfection complex formation and therefore should be excluded from the complex formation step. Transfection complexes can be added to cells grown in complete culture medium containing low levels of antibiotics (0.1 1X final concentration of penicillin/streptomycin mixture). Post-transfection incubation time. Determine the best incubation time post-transfection for each cell type. The optimal incubation time is generally hours, but will vary depending on the goal of the experiment, nature of the plasmid, and the half-life of the expressed protein. Do not use DNA prepared using miniprep kits for transfection. Do not use serum or antibiotics in the medium during transfection complex formation. Page 2 of 7
3 DNA TRANSFECTION PROTOCOL The following procedure describes how to perform DNA transfections in 6-well s. The surface areas of other culture vessels are different and transfections must be scaled accordingly. Appropriately increase or decrease the amounts of serum free medium, TransIT-Prostate Reagent, Prostate Boost Reagent, DNA and complete culture medium based on the surface area of the cell culture vessel. Table 1 presents recommended starting conditions depending on culture vessel size. Table 1. Recommended starting conditions for DNA transfections with the TransIT-Prostate Transfection Kit. Culture vessel 96-well 48-well 24-well 12-well 6-well 10-cm dish T75 flask Surface area 0.35 cm cm cm cm cm 2 59 cm 2 75 cm 2 Complete growth medium 92 µl 263 µl 0.5 ml 1 ml 2.5 ml 15.5 ml 19.7 ml Serum-free medium 9 µl 26 µl 50 µl 100 µl 250 µl 1.5 ml 1.9 ml DNA (1µg/µl stock) 0.09 µl 0.25 µl 0.5 µl 1 µl 2.5 µl 15.5 µl 19.7 µl TransIT-Prostate Reagent Prostate Boost Reagent* 0.18 µl 0.5 µl 1 µl 2 µl 5 µl 31 µl 39.4 µl 0.18 µl 0.5 µl 1 µl 2 µl 5 µl 31 µl 39.4 µl *Different Prostate Boost Reagent amounts are optimal for different prostate cell types. It may be necessary to titrate the Prostate Boost Reagent from 0 5 µl per 1 µg of DNA. Transient DNA transfection protocol per well of a 6-well A. Plate cells 1. Approximately hours before transfection, cells in 2.5 ml complete growth medium per well in a 6-well. Ideally cells should be 50 70% confluent (~ cells/well) prior to transfection. 2. Incubate the cell cultures overnight. B. Prepare TransIT-Prostate:Prostate Boost:DNA complexes (Immediately before transfection) 1. Warm TransIT-Prostate and Prostate Boost reagents to room temperature and vortex gently before using. 2. Place 250 µl of Opti-MEM I Reduced-Serum Medium in a sterile tube. 3. Add 2.5 µg (2.5 µl of a 1 µg/µl stock) DNA. Pipet gently to mix completely. 4. Add 5.0 µl TransIT-Prostate Reagent to the diluted DNA mixture. Pipet gently to mix completely. 5. Incubate at room temperature for 5 20 minutes. 6. Add 5.0 µl Prostate Boost Reagent to the diluted DNA mixture. Pipet gently to mix completely. 7. Incubate at room temperature for 5 20 minutes. Surface areas are based on Greiner tissue culture s and Falcon 10-cm dishes and T75 flasks. All volumes given are per well (or per dish) for a given culture vessel. If small volumes of reagents need to be pipetted, dilute the TransIT-Prostate and Prostate Boost reagent in 80% and 100% ethanol, respectively. Do not store diluted reagents. Divide cultured cells hours before transfection to ensure active cell division at the time of transfection. For loosely adherent cell types, e.g., LNCaP, use poly-lysine coated tissue culture s to aid in cell adherence. Warm TransIT-Prostate and Prostate Boost reagents to room temperature and vortex gently before each use. Page 3 of 7
4 C. Distribute the complexes to cells in complete growth medium 1. Add the complexes drop-wise to different areas of the wells. It is not necessary to replace the complete growth medium with fresh medium. 2. Gently rock the culture vessel back-and-forth and from side-to-side to evenly distribute the TransIT-Prostate:Prostate Boost:DNA complexes. 3. Incubate for hours. It is not necessary to replace the complete growth medium with fresh medium. 4. Harvest cells and assay as required. There is no need to change fresh culture medium after transfection. If required, perform a medium change at least 4 hours post-transfection. For generating stable cell transfectants, passage the cells hours post-transfection in complete growth medium containing the appropriate selection antibiotic such as G418 or Hygromycin B. Maintain selection for 1 2 weeks, allowing selection of cells that have undergone stable integration of DNA. Page 4 of 7
5 TROUBLESHOOTING GUIDE Problem Solution LOW DNA TRANSFECTION EFFICIENCY TransIT-Prostate or Prostate Boost Reagent was not mixed properly Suboptimal amount of TransIT-Prostate Reagent Suboptimal amount of Prostate Boost Reagent Warm TransIT-Prostate and Prostate Boost Reagents to room temperature and vortex gently before each use. Determine optimal amount of TransIT-Prostate Reagent for each Prostate cell type. Titrate the TransIT-Prostate Reagent from 2 5 µl per 1 µg DNA. Refer to Before You Start on Page 2. Determine optimal amount of Prostate Boost Reagent for each Prostate cell type. Titrate the Prostate Boost Reagent from 0 5 µl per 1 µg DNA. Refer to Before You Start on Page 2. Suboptimal DNA concentration Low-quality plasmid DNA Inhibitor present during transfection Incorrect vector sequence Confirm DNA concentration and purity. Use plasmid DNA preps that have an A 260/280 absorbance ratio of The optimal DNA concentration generally ranges between 1 3 µg/well of a 6-well. Start with 2.5 µg/well of a 6-well. Consider testing more or less DNA while scaling the amount of TransIT-Prostate and Prostate Boost Reagents accordingly. Use highly purified, sterile, endotoxin and contaminant-free DNA for transfection. We recommend using Mirus Bio s MiraCLEAN Endotoxin Removal Kit (MIR 5900) for removal of endotoxin from your DNA preparation. Alternatively, use cesium chloride gradient or anion exchange purified DNA which contains level of endotoxin that do not harm most cells. Do not use DNA prepared using miniprep kits as it may contain high levels of endotoxin. Serum and antibiotics inhibit transfection complex formation. Prepare transfection complexes in serum-free growth medium. We recommend Opti-MEM I Reduced-Serum Medium. Once transfection complexes are formed, they can be added directly to cells cultured in complete growth medium containing serum and 0.1 1X antibiotics. Polyanions such as dextran sulfate or heparin can inhibit transfection. Use culture medium that does not contain these polyanions. If necessary, the transfection medium can be replaced with polyanion containing medium 24 hours post transfection. If you do not observe expression of your target insert, verify the sequence of the plasmid DNA. Transfection incubation time Determine the optimal transfection incubation time for each cell type and experiment. Test a range of incubation times (e.g hours). The best incubation time is generally hours. Cells not actively dividing at the time of transfection Precipitate formation during transfection complex formation Divide the culture at least hours before transfection to ensure that the cells are actively dividing and reach optimal cell density at time of transfection. During complex formation, scale all reagents including serum-free medium, TransIT-Prostate Reagent, Prostate Boost Reagent and plasmid DNA according to Table 1 on Page 3. Precipitation may be observed when excess DNA is used during complex formation. This may negatively impact transfection efficiency. To avoid precipitation when using high concentrations of DNA, increase the volume of serum-free medium during complex formation by two-fold. Page 5 of 7
6 TROUBLESHOOTING GUIDE continued Problem Solution LOW DNA TRANSFECTION EFFICIENCY Proper experimental controls were not included HIGH CELLULAR TOXICITY Transfection complexes and cells not mixed thoroughly after complex addition Transfection complexes added to cells cultured in serum-free medium Endotoxin-contaminated plasmid DNA Expressed target gene is toxic to cells Cell density not optimal at time of transfection Cell morphology has changed To verify efficient transfection, use TransIT-Prostate Transfection Kit to deliver a positive control such as a luciferase, beta-galactosidase or green fluorescent protein (GFP) encoding plasmid. To assess delivery efficiency of plasmid DNA, use Mirus Label IT Tracker Intracellular Nucleic Acid Localization Kit to label the target plasmid or Mirus prelabeled Label IT Plasmid Delivery Controls (please refer to Related Products on Page 7). Add transfection complexes drop-wise to different areas of the wells containing d cells. Gently rock the dish back-and-forth and from side-to-side to distribute the complexes evenly. Do not swirl or rotate the dish, as this may cause uneven distribution. Allow transfection complexes to form in serum-free medium, then add these complexes to cells cultured in complete growth medium. The presence of serum in the growth medium improves transfection efficiency and reduces cytotoxicity. No culture medium change is required after the addition of transfection complexes to cells. Use highly purified, sterile, endotoxin and contaminant-free DNA for transfection. We recommend using Mirus Bio s MiraCLEAN Endotoxin Removal Kit (MIR 5900) for removal of any traces of endotoxin from your DNA preparation. Alternatively, use cesium chloride gradient or anion exchange purified DNA which contains levels of endotoxin that do not harm most cells. Do not use DNA prepared using miniprep kits as it might contain high levels of endotoxin. Compare toxicity levels against a cells alone control and cells transfected with an empty vector to assess the cytotoxic effects of the target protein being expressed. If lower levels of target gene expression are desired in your transfection experiments, consider reducing the amount of target plasmid. Maintain the optimal TransIT-Prostate:Prostate Boost:DNA ratio by using carrier DNA such as an empty cloning vector. Determine the best cell density for each Prostate cell type to maximize transfection efficiency. Use this cell density in subsequent experiments to ensure reproducibility. For most Prostate cell types, 50 70% confluence is recommended at transfection, but use of higher or lower densities may increase cell viability depending on cell type. Mycoplasma contamination can alter cell morphology and affect transfection efficiency. Check your cells for Mycoplasma contamination. Use a fresh frozen stock of cells or use appropriate antibiotics to eliminate Mycoplasma. A high or low cell passage number can make cells more sensitive and refractory to transfection. Maintain a similar passage number between experiments to ensure reproducibility. Page 6 of 7
7 RELATED PRODUCTS Ingenio Electroporation Solution and Kits Label IT Plasmid Delivery Controls Label IT Tracker Intracellular Nucleic Acid Localization Kits MiraCLEAN Endotoxin Removal Kits TransIT -3D Transfection Reagent TransIT Transfection Reagent TransIT-PRO Transfection Kit TransIT -LT1 Transfection Reagent TransIT Cell Line Specific Transfection Reagents and Kits TransIT -QR and TransIT -EE Delivery Solutions and Kits Reagent Agent Reagent Agent is an online tool designed to help determine the best solution for nucleic acid delivery based on in-house data, customer feedback and citations. Learn more at: For details on our products, visit or Contact Mirus Bio for additional information. Mirus Bio LLC 545 Science Drive Madison, WI Toll-free: Direct: Fax: All rights reserved. Mirus Bio LLC. All trademarks are the property of their respective owners. HeLaMONSTER, Ingenio, Label IT, MiraCLEAN, µarray, plive, siquest, TransIT, TransIT-Neural, TransIT-PRO and TransIT-TKO are registered trademarks of Mirus Bio LLC. Tracker is a trademark of Mirus Bio LLC. Reagent Agent is a service mark of Mirus Bio LLC. Use of Mirus Bio TransIT polyamine transfection reagents are covered by U.S. Patent No. 5,744,335, No. 6,180,784, No. 7,101,995, No. 7,601,367 and patents pending. The use of certain Mirus Bio transfection products are the subject o f one or more of U.S. Patents No. 7,335,509, No. 7,655,468 and/or other pending U.S. patent applications. Mirus Bio Label IT nucleic acid labeling and modifying reagents are covered by U.S. Patent No. 6,262,252, No. 6,593,465, No. 7,049,142, No. 7,326,780 and No. 7,491,538. Cy 3 and Cy 5 products or portions thereof are manufactured under license from Carnegie Mellon University and are covered by U.S. Patent No. 5,268,486. This product is sold to the Buyer with a limited license for Research Use Only and is not for clinical, therapeutic or diagnostic use in humans or animals. This product, or parts from this product, may not be re-packaged or re-sold without written permission from Mirus Bio LLC. The buyer agrees not to infringe upon Mirus patents or to attempt to reverse engineer, reconstruct, synthesize or otherwise modify Mirus Bio products. A license from Mirus Bio LLC is required for commercial application of this product. For obtaining a license to use this product for commercial application, contact Mirus Bio LLC, 545 Science Drive, Madison, WI license@mirusbio.com. For full terms and conditions, visit For publications citing the use of TransIT-Prostate Transfection Kit, visit ML026-Rev.E 0113 Page 7 of 7
TransIT -CHO Transfection Kit
INTRODUCTION TransIT -CHO Transfection Kit is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in CHO cells and cell types of associated lineage. TransIT-CHO Transfection
More informationTransIT -Keratinocyte Transfection Reagent
TransIT -Keratinocyte Transfection Reagent INTRODUCTION TransIT -Keratinocyte Transfection Reagent is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in keratinocyte
More informationTransIT -CHO Transfection Kit
INTRODUCTION TransIT -CHO Transfection Kit is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in CHO cells and cell types of associated lineage. TransIT-CHO Transfection
More informationTransIT -293 Transfection Reagent
INTRODUCTION TransIT -293 Transfection Reagent is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in HEK 293 and cell types of associated lineage. TransIT-293 provides
More informationTransfection Kit Protocol for MIR 2900, 2904, 2905, 2906
TransIT-HeLaMONSTER Transfection Kit INTRODUCTION TransIT-HeLaMONSTER Transfection Kit is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in HeLa cells and cell types
More informationTransIT -LT1 Transfection Reagent
INTRODUCTION TransIT -LT1 Transfection Reagent is a broad spectrum reagent that provides high efficiency plasmid DNA delivery in many mammalian cell types including primary cells. TransIT-LT1 is a low
More informationTransIT Transfection Reagent
INTRODUCTION TransIT -2020 Transfection Reagent is a high-performance, animal-origin free, broad spectrum reagent that provides exceptional transfection of plasmid DNA into mammalian cells. TransIT-2020
More informationTransIT -Jurkat Transfection Reagent
INTRODUCTION TransIT -Jurkat Transfection Reagent is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in Jurkat cells and cell types of associated lineage. Jurkat cells
More informationTransIT -BrCa Transfection Reagent
INTRODUCTION TransIT -BrCa Transfection Reagent is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in breast cancer and related cell types including: MCF-7, MDA-MB-231
More informationTransIT -Keratinocyte Transfection Reagent
TransIT -Keratinocyte Transfection Reagent Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2800 INTRODUCTION TransIT -Keratinocyte Transfection Reagent is specifically
More informationTransIT -CHO Transfection Kit
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2170 INTRODUCTION TransIT -CHO Transfection Kit is specifically optimized to provide exceptional transfection efficiency
More informationTransIT -293 Transfection Reagent
TransIT -293 Transfection Reagent Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2700 INTRODUCTION TransIT -293 Transfection Reagent is specifically optimized to provide
More informationTransfection Reagent INTRODUCTION SPECIFICATIONS MATERIALS. For Research Use Only. Materials Supplied. Materials required, but not supplied
TransIT-Neural Transfection Reagent INTRODUCTION TransIT-Neural Transfection Reagent is specifically optimized to provide exceptional transfection efficiency of plasmid DNA in various neural cell types.
More informationTransIT -Insect Transfection Reagent
INTRODUCTION TransIT -Insect Transfection Reagent is a novel transfection formulation specifically developed for high-performance transfection of plasmid DNA into insect cells such as: Sf9, High Five (BTI-TN-5B1-4),
More informationTransIT -BrCa Transfection Reagent
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/5500 INTRODUCTION TransIT -BrCa Transfection Reagent is specifically optimized to provide exceptional transfection efficiency
More informationTransIT Transfection Reagent
INTRODUCTION TransIT -2020 is a broad spectrum transfection reagent that provides superior transfection of plasmid DNA into mammalian cells. TransIT-2020 is suitable for both transient and stable transfection
More informationTransIT -Jurkat Transfection Reagent
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2120 INTRODUCTION TransIT -Jurkat Transfection Reagent is specifically optimized to provide exceptional transfection
More informationTransIT -LT1 Transfection Reagent
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2300 INTRODUCTION TransIT -LT1 Transfection Reagent is a broad spectrum reagent that provides high efficiency plasmid
More informationTransIT-TKO Transfection Reagent
INTRODUCTION TransIT-TKO is a broad spectrum sirna transfection reagent that enables high efficiency sirna delivery and knockdown of target gene expression in many cell types including primary cells. TransIT-TKO
More information3D Transfection System
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/5800 INTRODUCTION The 3D Transfection System containing the TransIT -3D Transfection Reagent and alvetex 12-well tissue
More informationTransIT-X2 Dynamic Delivery System
INTRODUCTION TransIT-X2 Dynamic Delivery System is an advanced, non-liposomal polymeric system that enables high efficiency transfection of many cell types, including primary cells. TransIT-X2 can be used
More informationTransIT -mrna Transfection Kit
INTRODUCTION TransIT -mrna Transfection Kit is designed to transfect RNA into a broad range of cell types with minimal cellular toxicity. RNA delivery avoids transcriptional regulation effects by directly
More informationTransIT -Insect Transfection Reagent
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/6100 INTRODUCTION TransIT -Insect Transfection Reagent is a novel transfection formulation specifically developed for
More informationTransIT-TKO Transfection Reagent
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2150 INTRODUCTION TransIT-TKO is a broad spectrum sirna transfection reagent that enables high efficiency sirna delivery
More informationTransfection Reagent Protocol for MIR 2110, 2114, 2115, 2116
TransIT-siQUEST Transfection Reagent INTRODUCTION TransIT-siQUEST is a broad spectrum sirna transfection reagent that enables high efficiency sirna delivery and knockdown of target gene expression in many
More informationTransIT -LT1 Transfection Reagent
Protocol for Product Nos. MIR 2300, 2304, 2305, 2306 i n t r o d u c t i o n The easy to use TransIT-LT1 (Low Toxicity) Transfection Reagent provides superior transfection efficiency, cell viability, and
More informationTransIT-PRO Transfection Kit Protocol for MIR 5700 and 5760
INTRODUCTION TransIT-PRO Transfection Kit consists of a DNA transfection reagent and boost combination specifically developed for mammalian protein production in suspension 293 and CHO derived cells. The
More informationTransfection Reagent SPECIFICATIONS MATERIALS. For Research Use Only. Materials Supplied. Materials required, but not supplied
TransIT-siQUEST Transfection Reagent Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2110 INTRODUCTION TransIT-siQUEST is a broad spectrum sirna transfection reagent
More informationTransIT -mrna Transfection Kit
Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/2225 INTRODUCTION TransIT -mrna Transfection Kit is designed to transfect RNA into a broad range of cell types with
More informationTransIT-PRO Transfection Kit Protocol for MIR 5700 and 5760
Quick Reference Protocol, SDS and Certificate of Analysis available at mirusbio.com/5700 INTRODUCTION TransIT-PRO Transfection Kit was specifically developed for mammalian protein production in suspension
More informationTransIT-PRO Transfection Reagent Protocol for MIR 5740 and 5750
Quick Reference Protocol, SDS and Certificate of Analysis available at mirusbio.com/5740 INTRODUCTION TransIT-PRO Transfection Reagent was developed by empirically testing proprietary lipid and polymer
More informationCHOgro Expression System
SDS and Certificate of Analysis available at mirusbio.com/6260 INTRODUCTION The CHOgro Expression System is an optimized platform for transient, high titer protein production in suspension CHO derived
More informationTransIT -Lenti Transfection Reagent
Quick Reference Protocol, SDS and Certificate of Analysis available at mirusbio.com/6600 INTRODUCTION Lentivirus is an enveloped, single-stranded RNA virus from the Retroviridae family capable of infecting
More informationM X 500 µl. M X 1000 µl
GeneGlide TM sirna Transfection Reagent (Catalog # M1081-300, -500, -1000; Store at 4 C) I. Introduction: BioVision s GeneGlide TM sirna Transfection reagent is a cationic proprietary polymer/lipid formulation,
More informationIngenio Electroporation Kits and Solution
Protocol for MIR 0111, 0112, 0113, 0114, 011, 0116, 0117, 0118, 0119 INTRODUCTION Ingenio Electroporation Kits and Solution provide a universal, high efficiency, low toxicity solution for electroporation
More informationTransIT -Lenti Transfection Reagent
Quick Reference Protocol, SDS and Certificate of Analysis available at mirusbio.com/6600 INTRODUCTION Lentivirus is an enveloped, single-stranded RNA virus from the Retroviridae family capable of infecting
More informationIngenio Electroporation Kits and Solution
Protocol for MIR 08, 09, 01, 0111, 0112, 0113, 0114, 011, 0116, 0117, 0118, 0119 Quick Reference Protocol, SDS and Certificate of Analysis available at mirusbio.com/0111 INTRODUCTION Ingenio Electroporation
More informationPlease read the entire protocol carefully and proceed with the section(s) appropriate for the desired experiment.
Page 1 of 9 Lit.# ML044 Rev.5/18/05 TransIT -sipak Trial Kit Product Components Quantity Product No. TransIT -sipak Trial Kit TransIT -siquest Reagent* TransIT-TKO Reagent* *To re-order components separately,
More informationProduct Quantity* Product No.
Page 1 of 6 Lit.# ML015 Rev.7/04 TransIT-TKO Transfection Reagent Product Quantity* Product No. TransIT-TKO Transfection Reagent 0.4 ml MIR 2154 1 ml MIR 2150 5 ml (5 x 1 ml) MIR 2155 10 ml (10 x 1 ml)
More informationAvalanche -Everyday Transfection Reagent
The Transfection & Gene Expression Experts Avalanche -Everyday Transfection Reagent Cat. No. EZT-EVDY-1 Description Size: 0.75 ml 1.5 ml Store at 4 C As the simplified version of our most popular and powerful
More informationOne tube (1.0 ml) TurboFectin 8.0 Transfection Reagent (80% ethanol solution)
Table of Contents Package Contents and Storage Conditions... 2 Related products... 2 Introduction... 3 Production and Quality Assurance:... 3 Experimental Procedures... 3 Transfection Optimization... 3
More informationAvalanche -Everyday Transfection Reagent
The Transfection & Gene Expression Experts Avalanche -Everyday Transfection Reagent Cat. No. EZT-EVDY-1 Description Size: 0.75 ml 1.5 ml Store at 4 C As the simplified version of our most popular and powerful
More informationAvalanche -Omni Transfection Reagent
The Transfection & Gene Expression Experts Avalanche -Omni Transfection Reagent Cat. No. EZT-OMNI-1 Size: 0.75 ml 1.5 ml Store at 4 C Description Avalanche -Omni Transfection Reagent is a proprietary lipid-polymer
More informationTransIT -VirusGEN Transfection Reagent
Quick Reference Protocol, SDS and Certificate of Analysis available at mirusbio.com/6700 INTRODUCTION Adeno-associated virus (AAV) is a nonenveloped, single stranded DNA virus from the Paroviridae family
More informationAvalanche -Omni Transfection Reagent
The Transfection & Gene Expression Experts Avalanche -Omni Transfection Reagent Cat. No. EZT-OMNI-1 Description Size: 0.75 ml 1.5 ml Store at 4 C Avalanche -Omni Transfection Reagent is a proprietary lipid-polymer
More informationPlease read the entire protocol carefully and proceed with the section(s) appropriate for the desired experiment.
Page 1 of 11 Lit.# ML045 Rev.5/18/05 TransIT -sipak Plus Trial Kit Product Components Component No. Quantity TransIT -siquest Reagent* 2111 0.1 ml Product No. TransIT -sipak Plus Trial Kit TransIT-TKO
More information293T/17 Cell Avalanche Transfection Reagent
The Transfection & Gene Expression Experts 293T/17 Cell Avalanche Transfection Reagent Cat. No. EZT-293T-1 Size: 0.5 ml 1.5 ml Store at 4 C Cell Line Information: Designations: 293T/17 [HEK 293T/17] Depositors:
More informationReagent. Cy 3 MIR Cy 5 MIR CX-Rhodamine MIR TM-Rhodamine MIR Biotin MIR 7204
Page 1 of 10 Lit.# ML023 Rev. 11/06 Label IT sirna Tracker Intracellular Localization Kit with TransIT-TKO Transfection Reagent Product Name Label IT sirna Tracker Reagent Product No. Cy 3 MIR 7200 Label
More informationPart Number: GY-SMPE010. Smart PEI. Instructions for Use. For research use only. Not for use in humans.
Transfection Part Number: GY-SMPE010 Reagent Instructions for Use For research use only. Not for use in humans. Transfection Reagent 1. Intended Use SmartPEI reagent is a low toxicity, serum-compatible
More informationMulti-Purpose Transfection Reagents
Product Information & Instruction Manual Multi-Purpose Transfection Reagents Cat. No: ScreenFect A S-3001 S-3001-2 S-3001-3 ScreenFect A-plus S-6001 S-6001-2 S-6001-3 www.incella.com Contents 1. Characteristics
More informationUser s Guide MegaTran 1.0 Transfection Reagent
User s Guide MegaTran 1.0 Transfection Reagent Package Contents and Storage Conditions... 2 Related products... 2 Introduction... 2 Production and Quality Assurance:... 3 Experimental Procedures... 3 Transfection
More informationPromoFectin-Hepatocyte Cell Transfection Reagent. Instruction Manual. Cat.No. PK-CT-2000-HEP-10 PK-CT-2000-HEP-50
PromoFectin-Hepatocyte Cell Transfection Reagent Instruction Manual Cat.No. PK-CT-2000-HEP-10 PK-CT-2000-HEP-50 2 Instruction Manual Contents Content 3 Formulation and Storage 3 General Considerations
More informationjetprime in vitro DNA & sirna transfection reagent PROTOCOL
jetprime in vitro DNA & sirna transfection reagent PROTOCOL DESCRIPTION jetprime is a novel powerful transfection reagent based on a polymer formulation manufactured at Polyplus-transfection. jetprime
More informationjetprime DNA & sirna transfection reagent
jetprime in vitro DNA & sirna Transfection Protocol Company Information... 2 Product Information... 3 1. Transient DNA transfection protocol... 4 1.1 Cell seeding... 4 1.2 DNA Transfection Protocol...
More informationflashbac Baculovirus Expression Systems flashbac, flashbac GOLD, flashbac ULTRA
flashbac Baculovirus Expression Systems Quick Reference Protocols, SDS, Product Data Sheets and Certificate of Analysis available at mirusbio.com/6115 INTRODUCTION The flashbac System is a streamlined
More informationGeneCellin TM Transfection Reagent Protocol
GeneCellin TM Transfection Reagent Protocol Table of content Description......3 Content......3 Storage......3 Certificate of quality......4 Parameters influencing transfection efficiency......4 Nucleic
More informationTHE DELIVERY EXPERTS PROTOCOL
THE DELIVERY EXPERTS jetprime in vitro DNA & sirna transfection reagent PROTOCOL DESCRIPTION jetprime is a novel powerful molecule based on a polymer formulation manufactured at Polyplustransfection. jetprime
More informationPromoFectin-HUVEC Cell Transfection Reagent. Instruction Manual. Cat.No. PK-CT-2000-HUV-10 PK-CT-2000-HUV-50
PromoFectin-HUVEC Cell Transfection Reagent Instruction Manual Cat.No. PK-CT-2000-HUV-10 PK-CT-2000-HUV-50 2 Instruction Manual Contents Content 3 Formulation and Storage 3 General Considerations 3 Transient
More informationingenio electroporation kits & solution
ingenio electroporation kits & solution Electroporation x DEFINITION and OPTIMIZATION What is ELECTROPORATION? Electroporation is a physical method of nucleic acid transfer wherein the cells and nucleic
More informationTransfection of Axol TM Human Neural Progenitor Cells (hnpcs) using Axol ReadyFect TM
Transfection of Axol TM Human Neural Progenitor Cells (hnpcs) using Axol ReadyFect TM Catalog No. ax0051 Application Protocol Version 2.0 Table of Contents Contents & Storage 3 Axol ReadyFect TM Technology
More informationSi Silencing Duplex. Advanced molecule designed for next-generation applications of RNAi.
SM Now improved with: Si 2 Purfect TRANSFECTION REAGENT PROTOCOL: 2 Si Silencing Duplex Advanced molecule designed for next-generation applications of RNAi. Greater Specificity of Suppression Reliable
More informationGeneCellin TM HTC Transfection Reagent Protocol vs6 2/12
GeneCellin TM HTC Transfection Reagent Protocol vs6 2/12 GeneCellin TM HTC Transfection Reagent Protocol Table of content Description......4 Content......4 Storage......4 Parameters influencing transfection
More informationPage 1 of 5. Product Name Label Quantity Product No. Cy 3 10 µg (~0.75 nmol) MIR Cy µg (~7.5 nmol) MIR 7901
Page 1 of 5 Label IT RNAi Delivery Control Product Name Label Quantity Product No. Cy 3 10 µg (~0.75 nmol) MIR 7900 Label IT RNAi Delivery Control Cy 3 100 µg (~7.5 nmol) MIR 7901 Fluorescein 10 µg (~0.75
More informationThis product is exclusively for research and for in-vitro application. It should not be used for therapeutic and diagnostic applications in humans.
AppliFect LowTox Transfection reagent for mammalian cells Product code A9027 Description AppliFect LowTox is a further development of existing reagents for liposome-mediated transfection of mammalian cells.
More informationTurboFect in vitro Transfection Reagent. Description. Storage. Reagents to be Supplied by the User. #R ml ( for in vitro transfections)
TurboFect in vitro Transfection Reagent #R0531 1 ml ( for 300-500 in vitro transfections) Description TurboFect in vitro Transfection Reagent* is a sterile solution of a cationic polymer in water. The
More informationThe highly efficient Transfection Reagent for Mammalian Cells. For ordering information, SDS, publications and application notes see
METAFECTENE PRO The highly efficient Transfection Reagent for Mammalian Cells For ordering information, SDS, publications and application notes see www.biontex.com Product Order No. Size METAFECTENE PRO
More informationFectoPRO DNA transfection kit for Bioproduction PROTOCOL
DNA transfection kit for Bioproduction PROTOCOL DESCRIPTION transfection kit is specifically designed for enhanced Transient Gene Expression using low DNA amounts, in suspension CHO and HEK-293 cells as
More informationTHE DELIVERY EXPERTS. INTERFERin in vitro sirna transfection reagent PROTOCOL
THE DELIVERY EXPERTS INTERFERin in vitro sirna transfection reagent PROTOCOL DESCRIPTION INTERFERin is a powerful sirna transfection reagent that ensures efficient gene silencing and reproducible transfection
More informationPromoFectin-siRNA. Instruction Manual
PromoFectin-siRNA 2 3 Contents Contents 3 Product Description 3 Materials and Methods 3 Troubleshooting 9 Shipping & Storage 9 Quality Control 9 Ordering Information 10 Contents 0.5 ml (PK-CT-2000-RNA-050),
More informationK2 Transfection System
K2 Transfection System DNA & RNA Transfection Kit for Mammalian Cells For order information, SDS, publications and application examples see www.biontex.com Product Order No. Size K2 Transfection System
More informationsirna Transfection Reagent
Description RiboJuice 0.3 ml 71115-3 1.0 ml 71115-4 Description RiboJuice efficiently delivers small interfering RNA (sirna) into a wide range of mammalian cell lines for targeted gene suppression (1).
More informationRiboCellin sirna Delivery Reagent Protocol vs1 2/12
RiboCellin sirna Delivery Reagent Protocol vs1 2/12 RiboCellin sirna Delivery Reagent Protocol Table of content Description......4 Content......4 Storage......4 Certificate of Quality......4 Parameters
More informationNucleic Acid Transfection. Genomics. TransIT Transfection Tools. Technical tip D.103
The nucleic acids delivery specialists design our transfection reagents. Our formulations provide superior transfection efficiency and cell viability. This complete range includes TransIT reagents, based
More informationThe RNAi Consortium (TRC) Broad Institute
TRC Laboratory Protocols Protocol Title: Lentivirus production of shrna, CRISPR, or ORF-pLX clones in 10 cm dishes or 6- well plates Current Revision Date: 6/3/2015 RNAi Platform,, trc_info@broadinstitute.org
More informationProtocol CRISPR Genome Editing In Cell Lines
Protocol CRISPR Genome Editing In Cell Lines Protocol 2: HDR donor plasmid applications (gene knockout, gene mutagenesis, gene tagging, Safe Harbor ORF knock-in) Notes: 1. sgrna validation: GeneCopoeia
More informationViaFect Transfection Reagent
TECHNICAL MANUAL ViaFect Transfection Reagent Instructions for Use of Products E4981, E4982 and E4983 Revised 7/17 TM409 ViaFect Transfection Reagent All technical literature is available at: www.promega.com/protocols/
More informationNanoFect Transfection Reagent Handbook
January 2008 NanoFect Transfection Reagent Handbook For efficient DNA transfection of a broad range of cell lines Sample & Assay Technologies QIAGEN Sample and Assay Technologies QIAGEN is the leading
More informationINTERFERin sirna transfection reagent
INTERFERin in vitro sirna Transfection Protocol Company Information... 2 Product Information... 3 1. Standard sirna transfection of adherent cells... 4 (forward transfection)... 4 1.1 Cell seeding... 5
More informationsitran TM sirna Transfection Application Guide
sitran TM sirna Transfection Application Guide Package Contents... 1 Storage conditions... 1 Related OriGene Products... 1 Notice to purchaser... 1 Introduction... 2 Experimental Procedures... 3 Transfection
More informationThe RNAi Consortium (TRC) Broad Institute
TRC Laboratory Protocols Protocol Title: Lentivirus production of shrna or ORF-pLX clones Current Revision Date: 10/20/2012 RNAi Platform,, trc_info@broadinstitute.org Brief Description: This protocol
More informationFectoCHO Expression system DNA transfection kit for protein production PROTOCOL
FectoCHO Expression system DNA transfection kit for protein production DESCRIPTION PROTOCOL FectoCHO Expression system is a three-component kit that includes FectoPRO transfection reagent, FectoPRO Booster
More informationThe Transfection Reagent for visualizing Lipofection with DNA. For ordering information, SDS, publications and application notes see
METAFECTENE FluoR The Transfection Reagent for visualizing Lipofection with DNA For ordering information, SDS, publications and application notes see www.biontex.com Product Order No. Size METAFECTENE
More informationDOTAP. The established Transfection Reagent for Mammalian Cells. For ordering information, SDS, publications and application notes see
DOTAP The established Transfection Reagent for Mammalian Cells For ordering information, SDS, publications and application notes see www.biontex.com Product Order No. Size DOTAP T010 1.0 1.0 ml DOTAP T010
More informationContents Introduction 2. Storage and Stability.. 2. Kit Contents Important Notes sirna Transfection Protocols 3
Contents Contents... 1 Introduction 2 Storage and Stability.. 2 Kit Contents... 2 Important Notes... 2 sirna Transfection Protocols 3 Optimizing sirna Transfection..... 3 Plasmid DNA Transfection.... 4
More informationINTERFERin in vitro sirna/mirna transfection reagent PROTOCOL
INTERFERin in vitro sirna/mirna transfection reagent PROTOCOL DESCRIPTION INTERFERin is a powerful sirna/mirna transfection reagent that ensures efficient gene silencing and reproducible transfection in
More informationjetpei -Macrophage in vitro DNA transfection reagent PROTOCOL
jetpei - in vitro DNA transfection reagent PROTOCOL DESCRIPTION jetpei - allows DNA transfection of macrophages and macrophage-like cells. It contains a mannose-conjugated linear polyethylenimine that
More informationFuGENE HD Transfection Reagent
For life science research only. Not for use in diagnostic procedures. FOR IN VITRO USE ONLY. FuGENE HD Transfection Reagent For the transient and stable transfection of animal and insect cells Cat. No.
More informationViaFect Transfection Reagent
TECHNICAL MANUAL ViaFect Transfection Reagent Instruc ons for Use of Products E4981, E4982 and E4983 1/14 TM409 ViaFect Transfection Reagent All technical literature is available at: www.promega.com/protocols/
More informationINSTRUCTION MANUAL HiViTM Transfection Reagent
Transfection reagents from labtech.com INSTRUCTION MANUAL HiViTM Transfection Reagent Labtech International Ltd 2 Birch House, Brambleside, Bellbrook Industrial Estate, Uckfield, TN22 1QQ Tel: 01825 744
More informationProduct: Arrest-In TM Transfection Reagent for RNAi
Product: Arrest-In TM Transfection Reagent for RNAi Catalog #: ATR1740, ATR1741, ATR1742, ATR1743 Product Description Arrest-In transfection reagent is a proprietary polymeric formulation, developed and
More informationjetpei -FluoF in vitro DNA Transfection Protocol
jetpei -FluoF in vitro DNA Transfection Protocol Company Information... 2 Product Information... 3 Contact our Technical Assistance and Scientific Advice Service... 4 1.Transfection protocol... 5 1. 1.
More informationFast and reliable purification of up to 100 µg of transfection-grade plasmid DNA using a spin-column.
INSTRUCTION MANUAL ZymoPURE Plasmid Miniprep Kit Catalog Nos. D4209, D4210, D4211 & D4212 (Patent Pending) Highlights Fast and reliable purification of up to 100 µg of transfection-grade plasmid DNA using
More informationIntroduction to Sinofection
Introduction to Sinofection Sinofection is a superior cationic polymer-based transfection reagent. It has been successfully applied to transfection at various scales over a broad range of cell lines. Compared
More informationCalPhos Mammalian Transfection Kit User Manual
CalPhos Mammalian Transfection Kit User Manual Cat. No. 631312 PT3025-1 (PR3Z643) Published 22 December 2003 Table of Contents I. Introduction 3 II. List of Components 4 III. Additional Materials Required
More informationsiport Amine Transfection Agent
siport Amine Transfection Agent Polyamine-Based Transfection Agent Part Number AM4502, AM4503 A. Introduction siport Amine Polyamine-Based Transfection Agent is a proprietary blend of polyamines formulated
More informationOZ Biosciences / Protocol FlyFectin / / - 0 -
OZ Biosciences / Protocol FlyFectin / www.ozbiosciences.com / - 0 - FlyFectin Instruction Manual FlyFectin The only specifically designed and very efficient transfection reagent for insect cells List of
More informationScreenFect mrna Transfection Reagent INSTRUCTION MANUAL
l ScreenFect mrna Transfection Reagent INSTRUCTION MANUAL S-5001 Cat. No. S-5001-2 S-5001-3 Contents 1. Overview: How to use ScreenFect mrna 2 2. Advantages of mrna Transfection and ScreenFect mrna 3 3.
More informationThe fastest, simplest method for purifying the highest quality endonuclease-free plasmid DNA.
INSTRUCTION MANUAL Zyppy Plasmid Midiprep Kit Catalog Nos. D4025 & D4026 (Patent Pending) Highlights The fastest, simplest method for purifying the highest quality endonuclease-free plasmid DNA. Pellet-Free
More informationAmaxa 96-well Shuttle Protocol for Primary Cell Optimization
For use with plasmid DNA and/or sirna The Primary Cell Optimization Protocol enables you to optimize 96-well Shuttle Conditions for a primary cell of your choice using our Primary Cell Optimization 96-well
More informationRNP Transfection in WTCs
Version 1.0 2016 Allen Institute for Cell Science RNP Transfection in WTCs Required reagent list: Complete mtesr1 culture media, referred to in this protocol as simply mtesr1 : 400 ml basal media with
More information